Contents About Avid Nano Dynamic Light Scattering Products - - PowerPoint PPT Presentation

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Contents About Avid Nano Dynamic Light Scattering Products - - PowerPoint PPT Presentation

Jun 30, 2023 119 likes •443 views

Contents About Avid Nano Dynamic Light Scattering Products Accessories About Avid Nano Established in July 2009 Based in High Wycombe, UK Design & manufacture novel DLS DYNAMIC LIGHT SCATTERING Dynamic Light

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Contents

 About Avid Nano  Dynamic Light Scattering  Products  Accessories

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About Avid Nano

 Established in July 2009  Based in High Wycombe, UK  Design & manufacture novel DLS

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DYNAMIC LIGHT SCATTERING

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Dynamic Light Scattering

 Observe time dependent intensity fluctuations

  • f light to directly measure...

– Hydrodynamic radius of molecules/particles in solution / suspension

  • Intensity size distribution
  • Mass size distribution
  • Aggregation
  • Molecular weight can be estimated
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Let's describe this visually

Scattering Volume

Ø40µm

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I

time Scattering Volume

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I

time Scattering Volume

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I

time Scattering Volume

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I

time Scattering Volume

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I

time Scattering Volume

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I

time

 Looks like random

noise, but...

 ...small sizes diffuse

more quickly than large sizes, so...

 ...rate of change tells

us the mean particle size

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Log10 time (µs) Amplitude, G2

 Intensity pattern

produces a correlation function and diffusion constant, Dt.

 Calculate mean

hydrodynamic radius, (Rh) and polydispersity index (PdI) Typical measurement time, 30s

Dynamic Light Scattering

Correlation function gives diffusion constant, Dt

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Dynamic Light Scattering

Rh= KT 6 Dt

We can use the 'Stokes- Einstein' equation to easily calculate the average hydrodynamic radius, Rh

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Size Distributions

0.01 0.1 1 10 100 1000 0.2 0.4 0.6 0.8 1 1.2

By comparing the measured data to a series of artificial correlation data we can produce a size distribution

Measured data Artificial data Size (nm) Intensity

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1.0 10.0 1.0 10.0 100.0 1000.0

f(x) = 2.75 x^2.49

Molecular Weight Estimator

Common Gobular Proteins

Monomer Radius (nm) MW (kDa)

To estimate molecular weight, we use a globular model derived from a curve

  • f common proteins

Molecular Weight Model

2.75r

2.49

Mw ~

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The model works well for many proteins

Molecular Weight Model

Molecule Size (nm)

  • Est. MW

(kDa) MW (kDa) Insulin (pH 2) 1.4 5.8 5.8 Lysozyme 2.0 14.5 14.7 Insulin (pH 7) 2.7 32.6 34.2 BSA 3.6 67.0 66.8 Hexokinase 4.3 104 102

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Typical Results

Experiment name Intensity Distribution Distribution Table of Results Mass Distribution

  • Ave. Correlation

Function Mean Size and Polydispersity

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Example Data

Highly monodisperse protein solution producing good quality crystals Narrow intensity peak Low polydispersity index Strong light scattering intensity from high concentration sample Mean radius 3.1nm used for molecular weight estimate (42kDa actual)

Mean radius Mw Estimate

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Example Data

Mixture of certified standards High PdI indicates broad or multi- mode distribution Intensity peaks confirm bi-modal distribution. Light scatters proportionally to Rh^6 Mass distribution indicates the amount of 10nm material much greater than 100nm

Mean radius. High Pd. Index

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Example Data

Overlaid Distributions Examples of mono-modal and multi-modal data. In each example we see normal variability caused by scattering intensity variation The size variability very low in main protein peak (7.1nm)

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Dynamic Light Scattering

 Primary data

– Mean hydrodynamic radius – Polydispersity index

 Secondary Data

– Intensity size distribution – Mass size distribution – Molecular weight estimate

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Applications

 Protein purification  Aggregation  Quick molecular weight estimate  Micelle formation  Thermal denaturing  Colloids and nano-particles

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Key Strengths

 Speed – especially with disposable cuvettes  Incredible sensitivity to aggregation  Requires little a priori knowledge  Absolute measurement - no calibration  Maintenance free

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Products

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W130i

 Designed for the protein specialist.

– Unbeatable sensitivity (0.1mg/ml, 15kDa protein) – 5µl disposable cuvette (standard) – Temperature control (0-90°C) – Compatible with standard cuvettes

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BladeCell Disposable Cuvette

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BladeCell Disposable Cuvette

DLS has reputation for being very useful but a bit tedious at times

– Expensive quartz cuvettes – Cleaning required – Cross-contamination issues

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BladeCell Disposable Cuvette

DLS has reputation for being very useful but a bit tedious at times

– Expensive quartz cuvettes – Cleaning required – Cross-contamination issues

BladeCell cuvette solves problem

– Only 5µl – No cleaning or reference m'ments – much faster than quartz – Full sample recovery – Zero cross-contamination

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Future Developments

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SMART-NANO

Commercializing hyper-sensitive DLS based on known light scattering techniques. Up to 10x sensitivity increase over current DLS performance. Suited to measurements at very low concentrations and for very small molecules Final negotiations September 2011. 2 year project commencing end 2011.

Sensitive MeAsuRemenT, detection, and identification of engineered NANO particles in complex matrices

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Thank you for watching